NFkB, p65 (Rel A) pS529 antibody

Antigen: NFkB, p65 (Rel A) pS529

Clonality: Polyclonal

Host: Rabbit

Reactivity: Cow (Bovine), Human, Mouse (Murine), Rat (Rattus), Sheep (Ovine)

Application: ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN105287

Additional Information

Immunogen: NFkB p65 (Rel A) peptide corresponding to a region near phospho Serine 529 of the human protein conjugated to Keyhole Limpet Hemocyanin (KLH). Sequence information: PNGLLpSGDEDFC

Format: Liquid (sterile filtered)

Specificity: This product was prepared from monospecific antiserum by delipidation and defibrination. This phospho specific polyclonal antibody is specific for phosphorylated pS529 human p65. Reactivity with non-phosphorylated p65 is minimal. Cross reactivity with pS529 phosphorylated p65 from mouse, rat or other species has not been determined.

Application Details

Application Notes: Recommended Dilutions: For immunoblotting a 1:1,000 dilution is recommended. A 65 kDa band corresponding to human p65 is detected. HeLa cells or TNF inducible KBM-5 cells can be used as a positive control. For ELISA a 1:5,000 to 1:10,000 dilution is recommended. Researchers should determine optimal titers for other applications. Note: This phospho specific polyclonal antibody reacts human pS529 p65 and shows minimal reactivity by western blot with non-phosphorylated p65 and minimal reactivity by ELISA against the non-phosphorylated form of the immunizing peptide. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation.

Concentration: 90.0 mg/ml (by Refractometry)

Buffer: Stabilizer: None. Preservative: 0.1% (w/v) Sodium Azide. Buffer: None.

Storage: Store vial at -20° C prior to opening. Centrifuge product if not completely clear after standing atroom temperature. Dilute only prior to immediate use. For extended storage aliquot contents and freeze at -20° C orbelow. Avoid cycles of freezing and thawing. Expiration date is one (1) year from date of opening.

Research Area: Transcription Factors

Restrictions: For Research Use only

Images

TNF Induces phosphorylation of p65 in KBM-5 cells. Nuclear protein lysates prepared after 0, 5, 10, 15, 30 and 60 minutes of 0.1 nM TNF treatment of KBM-5 cells shows inducible phosphorylation using phospho specific polyclonal anti-human pS529 p65. Anti-beta-actin staining confirms loading of equivalent amounts of protein. HRP conjugated Gt-anti-Rabbit IgG was used to develop the blot using a chemiluminescent detection method. Other detection methods will yield similar results. Data contributed by Aggarwal BB, personal communication.

Publications

Publications: Shakibaei, John, Schulze-Tanzil et al.: "Suppression of NF-kappaB activation by curcumin leads to inhibition of expression of cyclo-oxygenase-2 and matrix metalloproteinase-9 in human articular chondrocytes: Implications for the treatment of osteoarthritis." in: Biochemical pharmacology, Vol. 73, Issue 9, pp. 1434-45, 2007 (PubMed).

Amith, Jayanth, Franchuk et al.: "Neu1 desialylation of sialyl alpha-2,3-linked beta-galactosyl residues of TOLL-like receptor 4 is essential for receptor activation and cellular signaling." in: Cellular signalling, 2009 (PubMed).

Gavert, Ben-Shmuel, Lemmon et al.: "Nuclear factor-kappaB signaling and ezrin are essential for L1-mediated metastasis of colon cancer cells." in: Journal of cell science, Vol. 123, Issue Pt 12, pp. 2135-43, 2010 (PubMed).